晶状体上皮细胞论文热休克预处理对人晶状体上皮细胞HSP60mRNA表达的影响及意义.doc

晶状体上皮细胞论文：热休克预处理对人晶状体上皮细胞HSP60mRNA表达的影响及意义 【中文摘要】采用SYBR Green I实时荧光定量PCR方法检测人晶状体上皮细胞在不同环境刺激处理后HSP60mRNA的表达的变化,探讨HSP60mRNA表达变化规律以及不同环境刺激处理后表达是否有差异。方法采用正常人组织源性晶状体上皮细胞HUM-CELL-0098进行细胞传代培养后,将传代培养后的细胞进行分组实验,在氧化条件以及热休克处理后加氧化条件下培养细胞30min后,在正常培养条件下继续培养并且选取不同时间点(Oh、2h、4h、6h、16h、24h)的细胞采用SYBR Green I实时荧光定量PCR检测HSP60mRNA的表达情况。结果观察到晶状体上皮细胞在不同环境刺激下均表达HSP60mRNA,但热休克预处理后的细胞HSP60mRNA的表达较未热休克处理细胞高。氧化应激条件下HSP60mRNA的表达明显增加,在刺激结束后表达含量为最高,以后逐渐下降；在热休克预处理组中HSP60mRNA的表达含量也明显增加,但在4h表达量最高,以后逐渐下降。两组细胞在不同时段HSP60mRNA表达量变化趋势不同F=178.786,P=0.007,P0.05,有统计学意义。此外,热休克预处理组与单纯氧化应激组中HSP60mRNA表达含量有显著差异,热休克处理组的表达含量较单纯氧化应激组高,存在组间差别F=193.118,P=0.001,P0.05,有统计学意义。时间与分表交互作用F=124.939,P=0.003,有统计学意义。两组细胞HSP60mRNA表达量进行各个时间两两对比发现这两组数据在统计学上有差异。此外,还发现热休克处理后的细胞表达HSP60mRNA可能具有时间依赖性。结论人晶状体上皮细胞在不同应激条件下,均能诱导HSP60mRNA的合成,但在不同应激条件下的HSP60mRNA的表达是具有差异的,热休克预处理后的细胞HSP60mRNA的表达高于氧化损伤处理的细胞,给予预先的热休克处理可通过提高HSP60mRNA表达来保护晶状体。 【英文摘要】Adopt SYBR GreenⅠreal-time quantitative polymerase chain reaction (PCR) testing to investigate the expression of HSP60mRNA of human lens epithelial cells in different environmental stimuli, discusses the change rule of HSP60 express quantity and the expression of HSP60mRNA in different environment whether is different.MethodsHuman lens epithelial cells HUM-CELL-0098 were training and then divided into groups. LECs were exposed in the oxidation conditions and high temperature add oxidation conditions during 30min and then recovered to physiological condition. At different interval (0 hour,2 hour,4 hour,6 hour,16 hour,24 hour), these groups were tested by SYBR GreenⅠreal-time fluorescence quantitative polymerase chain reaction (PCR).ResultsThe expression of HSP60 was existed both in the different environment, but cells that in oxidative stress condition HSP60mRNA expression less than cells in heat shock group. The level of HSP60 mRNA was increased soon after oxidative stress condition, and the level at 0 hour is highest, then expression level decreased gradually later; In the heat shock pretreatment gr